Compounds useful for inhibition of farnesyl protein transferase

ABSTRACT

Novel compounds of the formula: ##STR1## are disclosed. Compounds of Formula 1.0 are represented by the compounds of formulas: ##STR2## wherein R 1 , R 3  and R 4  are each independently selected from halo. Also disclosed are methods of inhibiting farnesyl protein transferase and the growth of abnormal cells, such as tumor cells.

REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application Ser.No. 60/026,114 filed Sep. 13, 1996, and U.S. Provisional ApplicationSer. No. 60/050,009 filed Jun. 17, 1997.

BACKGROUND

WO 95/10516, published Apr. 20, 1995 discloses tricyclic compoundsuseful for inhibiting farnesyl protein transferase.

In view of the current interest in inhibitors of farnesyl proteintransferase, a welcome contribution to the art would be compounds usefulfor the inhibition of farnesyl protein transferase. Such a contributionis provided by this invention.

SUMMARY OF THE INVENTION

This invention provides compounds useful for the inhibition of farnesylprotein transferase (FPT). The compounds of this invention arerepresented by the formula: ##STR3## or a pharmaceutically acceptablesalt or solvate thereof, wherein: one of a, b, c and d represents N orNR⁹ wherein R⁹ is O⁻, --CH₃ or --(CH₂)_(n) CO₂ H wherein n is 1 to 3,and the remaining a, b, c and d groups represent CR¹ or CR2; or

each of a, b, c, and d are independently selected from CR¹ or CR² ;

each R¹ and each R² is independently selected from H, halo, --CF₃,--OR¹⁰ (e.g., --OCH₃), --COR¹⁰, --SR¹⁰ (e.g., --SCH₃ and --SCH₂ C₆ H₅),--S(O)_(t) R¹¹ (wherein t is 0, 1 or 2, e.g., --SOCH₃ and --SO₂ CH₃),--SCN, --N(R¹⁰)₂, --NR¹⁰ R¹¹, --NO₂, --OC(O)R¹⁰, --CO₂ R¹⁰, --OCO₂ R¹¹,--CN, --NHC(O)R¹⁰, --NHSO₂ R¹⁰, --CONHR¹⁰, --CONHCH₂ CH₂ OH, --NR¹⁰COOR¹¹, ##STR4## --SR¹¹ C(O)OR¹¹ (e.g., --SCH₂ CO₂ CH₃), --SR¹¹ N(R⁷⁵)₂wherein each R⁷⁵ is independently selected from H and --C(O)OR¹¹ (e.g.,--S(CH₂)₂ NHC(O)O-t-butyl and --S(CH₂)₂ NH₂), benzotriazol-1-yloxy,tetrazol-5-ylthio, or substituted tetrazol-5-ylthio (e.g., alkylsubstituted tetrazol-5-ylthio such as 1-methyl-tetrazol-5-ylthio),alkynyl, alkenyl or alkyl, said alkyl or alkenyl group optionally beingsubstituted with halo, --OR¹⁰ or --CO₂ R¹⁰ ;

R³ and R⁴ are the same or different and each independently represents H,any of the substituents of R¹ and R², or R³ and R⁴ taken togetherrepresent a saturated or unsaturated C₅ -C₇ fused ring to the benzenering (Ring III);

R⁵, R⁶, R⁷ and R⁸ each independently represents H, --CF₃, --COR¹⁰, alkylor aryl, said alkyl or aryl optionally being substituted with --OR¹⁰,--SR¹⁰, --S(O)_(t) R¹¹, --NR¹⁰ COOR¹¹, --N(R¹⁰)₂, --NO₂, --COR¹⁰,--OCOR¹⁰, --OCO₂ R¹¹, --CO₂ R¹⁰, OPO₃ R¹⁰ or R⁵ is combined with R⁶ torepresent ═O or ═S and/or R⁷ is combined with R⁸ to represent ═O or ═S;

R¹⁰ represents H, alkyl, aryl, or aralkyl (e.g., benzyl);

R¹¹ represents alkyl or aryl;

X represents N, CH or C, which C may contain an optional double bond(represented by the dotted line) to carbon atom 11;

the dotted line between carbon atoms 5 and 6 represents an optionaldouble bond, such that when a double bond is present, A and Bindependently represent --R¹⁰, halo, --OR¹¹, --OCO₂ R¹¹ or --OC(O)R¹⁰,and when no double bond is present between carbon atoms 5 and 6, A and Beach independently represent H₂, --(OR¹¹)₂ ; H and halo, dihalo, alkyland H, (alkyl)₂, --H and --OC(O)R¹⁰, H and --OR¹⁰, ═O, aryl and H,═NOR¹⁰ or --O--(CH₂)_(p) --O-- wherein p is 2, 3 or 4; and

W represents a group selected from: ##STR5## wherein: R¹² is selectedfrom the group consisting of: (1) H; (2) alkyl (e.g., methyl and ethyl);(3) aryl; (4) arylalkyl (aralkyl);

R¹³ is selected from the group consisting of: (1) H; (2) alkyl (e.g.,methyl and ethyl); (3) alkoxy (e.g., methoxy); (4) heterocycloalkyl,e.g., (a) tetrahydopyranyl, and (b) substituted tetrahydropyranylwherein said substituents are selected from hydroxy and hydroxyalkyl(e.g., hydroxymethyl), for example D-galactosyl, i.e., ##STR6## (5)aryl; and (6) aralkyl, e.g., benzyl; R¹⁴ is selected from the groupconsisting of: (1) H; (2) alkyl (e.g., --C(CH₃)₃); (3) aryl; and (4)heteroaryl; ##STR7## represents a heterocycloalkyl ring wherein Yrepresents the remainder of the ring, said remainder comprising carbonatoms and optionally a hetero atom selected from the group consistingof: NH, NR¹⁵, O and S, and said remainder optionally having an aryl ring(e.g., phenyl) fused thereto; generally the heterocycloalkyl ringcontains 4 or 5 carbon atoms and usually 4 carbon atoms, examplesinclude: ##STR8## examples of a heterocycloalkyl ring having a aryl ringfused to the remainder Y include ##STR9## R¹⁵ represents --C(O)OR¹⁶ ;and R¹⁶ represents alkyl, preferably --C(CH₃)₃.

The compounds of this invention: (i) potently inhibit farnesyl proteintransferase, but not geranylgeranyl protein transferase I, in vitro;(ii) block the phenotypic change induced by a form of transforming Raswhich is a farnesyl acceptor but not by a form of transforming Rasengineered to be a geranylgeranyl acceptor; (iii) block intracellularprocessing of Ras which is a farnesyl acceptor but not of Ras engineeredto be a geranylgeranyl acceptor; and (iv) block abnormal cell growth inculture induced by transforming Ras.

The compounds of this invention inhibit farnesyl protein transferase andthe farnesylation of the oncogene protein Ras. Thus, this inventionfurther provides a method of inhibiting ras farnesyl proteintransferase, (e.g., ras farnesyl protein transferase) in mammals,especially humans, by the administration of an effective amount of thetricyclic compounds described above. The administration of the compoundsof this invention to patients, to inhibit farnesyl protein transferase,is useful in the treatment of the cancers described below.

This invention provides a method for inhibiting or treating the abnormalgrowth of cells, including transformed cells, by administering aneffective amount of a compound of this invention. Abnormal growth ofcells refers to cell growth independent of normal regulatory mechanisms(e.g., loss of contact inhibition). This includes the abnormal growthof: (1) tumor cells (tumors) expressing an activated Ras oncogene; (2)tumor cells in which the Ras protein is activated as a result ofoncogenic mutation in another gene; and (3) benign and malignant cellsof other proliferative diseases in which aberrant Ras activation occurs.

This invention also provides a method for inhibiting or treating tumorgrowth by administering an effective amount of the tricyclic compounds,described herein, to a mammal (e.g., a human) in need of such treatment.In particular, this invention provides a method for inhibiting ortreating the growth of tumors expressing an activated Ras oncogene bythe administration of an effective amount of the above describedcompounds. Examples of tumors which may be inhibited or treated include,but are not limited to, lung cancer (e.g., lung adenocarcinoma),pancreatic cancers (e.g., pancreatic carcinoma such as, for example,exocrine pancreatic carcinoma), colon cancers (e.g., colorectalcarcinomas, such as, for example, colon adenocarcinoma and colonadenoma), myeloid leukemias (for example, acute myelogenous leukemia(AML)), thyroid follicular cancer, myelodysplastic syndrome (MDS),bladder carcinoma, epidermal carcinoma, breast cancer and prostatecancer.

It is believed that this invention also provides a method for inhibitingor treating proliferative diseases, both benign and malignant, whereinRas proteins are aberrantly activated as a result of oncogenic mutationin other genes--i.e., the Ras gene itself is not activated by mutationto an oncogenic form--with said inhibition or treatment beingaccomplished by the administration of an effective amount of thetricyclic compounds described herein, to a mammal (e.g., a human) inneed of such treatment. For example, the benign proliferative disorderneurofibromatosis, or tumors in which Ras is activated due to mutationor overexpression of tyrosine kinase oncogenes (e.g., neu, src, abl,Ick, and fyn), may be inhibited or treated by the tricyclic compoundsdescribed herein.

The tricyclic compounds useful in the methods of this invention inhibitor treat the abnormal growth of cells. Without wishing to be bound bytheory, it is believed that these compounds may function through theinhibition of G-protein function, such as ras p21, by blocking G-proteinisoprenylation, thus making them useful in the treatment ofproliferative diseases such as tumor growth and cancer. Without wishingto be bound by theory, it is believed that these compounds inhibit rasfarnesyl protein transferase, and thus show antiproliferative activityagainst ras transformed cells.

DETAILED DESCRIPTION OF THE INVENTION

As used herein, the following terms are used as defined below unlessotherwise indicated:

MH⁺ -represents the molecular ion plus hydrogen of the molecule in themass spectrum;

benzotriazol-1-yloxy represents ##STR10##

1-methyl-tetrazol-5-ylthio represents ##STR11## alkenyl-representsstraight and branched carbon chains having at least one carbon to carbondouble bond and containing from 2 to 12 carbon atoms, preferably from 2to 6 carbon atoms and most preferably from 3 to 6 carbon atoms;

alkynyl-represents straight and branched carbon chains having at leastone carbon to carbon triple bond and containing from 2 to 12 carbonatoms, preferably from 2 to 6 carbon atoms;

alkyl-(including the alkyl portions of alkoxy, aralkyl andheteroarylalkyl)-represents straight and branched carbon chains andcontains from one to twenty carbon atoms, preferably one to six carbonatoms;

aralkyl-represents an aryl group, as defined below, bound to an alkylgroup, as defined above, preferably the alkyl group is --CH₂ --, (e.g.,benzyl);

aryl (including the aryl portion of aralkyl)-represents a carbocyclicgroup containing from 6 to 15 carbon atoms and having at least onearomatic ring (e.g., aryl is a phenyl ring), with all availablesubstitutable carbon atoms of the carbocyclic group being intended aspossible points of attachment, said carbocyclic group being optionallysubstituted (e.g., 1 to 3) with one or more of halo, alkyl, hydroxy,alkoxy, phenoxy, CF₃, amino, alkylamino, dialkylamino, --COOR¹⁰ or --NO₂;

--CH₂ -imidazolyl represents an imidazolyl group bound by anysubstitutable carbon of the imidazole ring to a --CH₂ --, that is:##STR12## such as --CH₂ --(2-, 4- or 5-)imidazolyl, for example##STR13## halo-represents fluoro, chloro, bromo and iodo;heteroaryl-represents cyclic groups, optionally substituted with R³ andR⁴, having at least one heteroatom selected from O, S or N, saidheteroatom interrupting a carbocyclic ring structure and having asufficient number of delocalized pi electrons to provide aromaticcharacter, with the aromatic heterocyclic groups preferably containingfrom 2 to 14 carbon atoms, e.g., (2-, 4- or 5-)imidazolyl, triazolyl,2-, 3- or 4-pyridyl or pyridyl N-oxide (optionally substituted with R³and R⁴), wherein pyridyl N-oxide can be represented as: ##STR14##heteroarylalkyl (heteroaralkyl)-represents a heteroaryl group, asdefined above, bound to an alkyl group, as defined above, preferably thealkyl group is --CH₂ -- (e.g., --CH₂ --(4- or 5-)imidazolyl);

heterocycloalkyl-represents a saturated, branched or unbranchedcarbocylic ring containing from 3 to 15 carbon atoms, preferably from 4to 6 carbon atoms, which carbocyclic ring is interrupted by 1 to 3hetero groups selected from --O--, --S-- or --NR¹⁰ --; suitableheterocycloalkyl groups include: (1) 2- or 3-tetrahydrofuranyl, (2) 2-or 3-tetrahydrothienyl, (3) 2-, 3- or 4-piperidinyl, (4) 2- or3-pyrrolidinyl, (5) 2- or 3-piperazinyl, (6) 2- or 4-dioxanyl, (7)tetrahydopyranyl, and (8) substituted tetrahydropyranyl wherein saidsubstituents are selected from hydroxy and hydroxyalkyl (e.g.,hydroxymethyl), for example, D-galactosyl, e.g., ##STR15##

The following solvents and reagents are referred to herein by theabbreviations indicated: ethanol (EtOH); methanol (MeOH); acetic acid(HOAc or AcOH); ethyl acetate (EtOAc); N,N-dimethylformamide (DMF);trifluoroacetic acid (TFA); trifluoroacetic anhydride (TFAA);1-hydroxybenzo-triazole (HOBT); 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (DEC); diisobutyl aluminum hydride (DIBAL);and 4-methylmorpholine (NMM).

Reference to the position of the substituents R¹, R², R³, and R⁴ isbased on the numbered ring structure: ##STR16##

Those skilled in the art will also appreciate that the S and Rstereochemistry at the C-11 bond are: ##STR17##

Compounds of Formula 1.0 include compounds wherein the bottompiperidinyl group is a 4- or 3-piperidinyl group, i.e., ##STR18##

Compounds of Formula 1.0 include compounds wherein R² and R⁴ are H, andR¹ and R³ are halo (preferably independently selected from Br or Cl).For example, R¹ is Br and R³ is Cl. These compounds include compoundswherein R¹ is in the 3-position and R³ is in the 8-position, e.g., 3-Brand 8-Cl. Compounds of Formula 1.0 also include compounds wherein R² isH, and R¹, R³ and R⁴ are halo (preferably independently selected from Bror Cl).

Preferably, compounds of Formula 1.0 are represented by compounds ofFormula 1.1: ##STR19## wherein all substituents are as defined forFormula 1.0.

Preferably, R² is H and R¹, R³ and R⁴ are halo; a is N and b, c and dare carbon; A and B are each H₂ ; the optional bond between C5 and C6 isabsent; X is CH; and R⁵, R⁶, R⁷ and R⁸ are H. More preferably, R¹, R³and R⁴ are independently selected from Br or Cl. Most preferably, R¹ isBr, and R³ and R⁴ are independently selected from Cl and Br.

More preferably, compounds of Formula 1.0 are represented by compoundsof Formula 1.2 and Formula 1.3: ##STR20## and most preferably, compoundsof Formulas 1.4 and 1.5 ##STR21## wherein R¹, R³ and R⁴ are eachindependently selected from halo, preferably, Br or Cl; and A, B, X andW are as defined for Formula 1.0. More preferably, A and B are each H₂ ;the optional bond between C5 and C6 is absent; and X is CH. Mostpreferably, R¹ is Br; R³ and R⁴ are independently Br or Cl, and stillmore preferably R³ is Cl and R⁴ is Br; A and B are each H₂ ; theoptional bond between C5 and C6 is absent; X is CH; and R⁵, R⁶, R⁷ andR⁸ are H.

When W represents: ##STR22## preferably R¹² is selected from the groupconsisting of: H, alkyl (most preferably methyl or ethyl) and aralkyl(most preferably benzyl); and preferably R¹³ is selected from the groupconsisting of: H, alkyl (most preferably methyl or ethyl), alkoxy (mostpreferably methoxy), aralkyl (most preferably benzyl), andheterocycloalkyl (most preferably D-galactosyl). Preferably: (1) R¹² andR¹³ are H; (2) R¹² is alkyl (e.g., methyl), and R¹³ is alkoxy (e.g.,methoxy); (3) R¹² is alkyl and R¹³ is alkyl (e.g, both R¹² and R¹³ areethyl); (4) R¹² is heterocycloalkyl (e.g., D-galactosyl) and R¹³ is H;or (5) R¹² is aralkyl (e.g., benzyl) and R¹³ is H.

When W represents ##STR23## the heterocycloalkyl ring ##STR24##preferably is selected from the group consisting of: ##STR25##

When W represents ##STR26## R¹⁴ is preferably alkyl and most preferably--C(CH₃)₃.

Compounds of Formulas 1.2A and 1.3A: ##STR27## are preferred when X isCH or N, and R¹, R³ and R⁴ are halo.

The preferred compounds of this invention are represented by thecompounds of Formulas: ##STR28## wherein R¹, R³ and R⁴ are halo and theremaining substituents are as defined above, with the compounds ofFormula 1.5A being more preferred.

Representative compounds of Formula 1.0 wherein W is ##STR29## include:##STR30##

Representative compounds of Formula 1.0 wherein W is ##STR31## include:##STR32##

Representative compounds of Formula 1.0 wherein W is ##STR33## include:##STR34##

Representative compounds of Formula 1.0 include: ##STR35##

The compounds of this invention also include the 1-N-oxides--i.e, forexample, compounds of the the formula: ##STR36## wherein represents theremainder of the compound, or pharmaceutically acceptable salts orsolvates thereof.

Optical rotation of the compounds ((+)- or (-)-) are measured inmethanol or ethanol at 25° C.

This invention includes the above compounds in the amorphous state or inthe cyrstalline state.

Lines drawn into the ring systems indicate that the indicated bond maybe attached to any of the substitutable ring carbon atoms.

Certain compounds of the present invention may exist in differentisomeric forms (e.g., enantiomers or diastereoisomers) includingatropisomers (i.e., compounds wherein the 7-membered ring is in a fixedconformation such that the 11-carbon atom is positioned above or belowthe plane of the fused beznene rings due to the presence of a 10-bromosubstituent). The invention contemplates all such isomers both in pureform and in admixture, including racemic mixtures. Enol forms are alsoincluded.

Certain tricyclic compounds will be acidic in nature, e.g. thosecompounds which possess a carboxyl or phenolic hydroxyl group. Thesecompounds may form pharmaceutically acceptable salts. Examples of suchsalts may include sodium, potassium, calcium, aluminum, gold and silversalts. Also contemplated are salts formed with pharmaceuticallyacceptable amines such as ammonia, alkyl amines, hydroxyalkylamines,N-methylglucamine and the like.

Certain basic tricyclic compounds also form pharmaceutically acceptablesalts, e.g., acid addition salts. For example, the pyrido-nitrogen atomsmay form salts with strong acid, while compounds having basicsubstituents such as amino groups also form salts with weaker acids.Examples of suitable acids for salt formation are hydrochloric,sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic,fumaric, succinic, ascorbic, maleic, methanesulfonic and other mineraland carboxylic acids well known to those in the art. The salts areprepared by contacting the free base form with a sufficient amount ofthe desired acid to produce a salt in the conventional manner. The freebase forms may be regenerated by treating the salt with a suitabledilute aqueous base solution such as dilute aqueous NaOH, potassiumcarbonate, ammonia and sodium bicarbonate. The free base forms differfrom their respective salt forms somewhat in certain physicalproperties, such as solubility in polar solvents, but the acid and basesalts are otherwise equivalent to their respective free base forms forpurposes of the invention.

All such acid and base salts are intended to be pharmaceuticallyacceptable salts within the scope of the invention and all acid and basesalts are considered equivalent to the free forms of the correspondingcompounds for purposes of the invention.

Compounds of the invention may be prepared according to the proceduresdescribed in WO 95/10516 published Apr. 20, 1995, application Ser. No.08/410,187 filed Mar. 24, 1995, application Ser. No. 08/577,951 filedDec. 22, 1995 (now abandoned), application Ser. No. 08/615,760 filedMar. 13, 1996 (now abandoned), WO 97/23478 published Jul. 3, 1997 whichdiscloses the subject matter of Ser. Nos. 08/577,951 and 08/615,760,application Ser. No. 08/711,925 filed Sep. 13, 1996, and applicationSer. No. 08/877,453 filed Jun. 17, 1997; the disclosures of each beingincorporated herein by reference thereto; and according to theprocedures described below.

Compounds of the invention can be prepared by reacting a compound of theformula: A ##STR37## wherein all substituents are as defined for Formula1.0, with the appropriate protected piperidinyl acetic acid (e.g.,1-N-t-butoxy-carbonylpiperidinyl acetic acid together with DEC/HOBT/NMMin DMF at about 25° C. for about 18 hours to produce a compound of theformula: ##STR38## The compound of Formula 13.0 is then reacted eitherwith TFA or 10% sulfuric acid in dioxane and methanol followed by NaOHto produce the compound of Formula 14.0 ##STR39##

For example, the compound of formula ##STR40## can be prepared byreaction of a compound of Formula 12.0 with1-N-t-butoxycarbonylpiperidinyl-4-acetic acid as described above.

For Example, compounds of Formula 15.0 include the compounds: ##STR41##The preparation of these compounds are described in Preparative Examples3, 4, 5, 6, 7, 8, 9, 10, 11, 12 and 13, respectively, below.

The compounds of the invention can be prepared by reacting a compound ofthe formula: ##STR42## with the appropriate protected piperidinyl aceticacid (e.g., 1-N-t-butoxycarbonylpiperidinyl acetic acid together withDEC/HOBT/NMM in DMF at about 25° C. for about 18 hours to produce acompound of the formula: ##STR43## The compound of Formula 13.1 is thenreacted either with TFA or 10% sulfuric acid in dioxane and methanolfollowed by NaOH to produce the compound of Formula 15.1 ##STR44##

The amide compounds of this invention, represented by Formula 1.7##STR45## can be prepared by reacting the compound of Formula 15.1 withthe appropriate carboxylic acid in the presence of a coupling agent suchas DEC and HOBT in dimethylformamide. Alternatively, the compound ofFormula 15.1 can be reacted with an acid chloride or anhydride in asolvent such as pyridine.

Compounds having an 1-N--O group: ##STR46## can be prepared from thecorresponding pyridyl compounds: ##STR47## by oxidation withmeta-chloroperoxybenzoic acid. This reaction is conducted in a suitableorganic solvent, e.g., dichloromethane (usually anhydrous) or methylenechloride, at a suitable temperature, to produce the compounds of theinvention having the N--O substituent at position 1 of Ring I of thetricyclic ring system.

Generally, the organic solvent solution of the starting tricyclicreactant is cooled to about 0° C. before the m-chloroper-oxybenzoic acidis added. The reaction is then allowed to warm to room temperatureduring the reaction period. The desired product can be recovered bystandard separation means. For example, the reaction mixture can bewashed with an aqueous solution of a suitable base, e.g., saturatedsodium bicarbonate or NaOH (e.g., 1N NaOH), and then dried overanhydrous magnesium sulfate. The solution containing the product can beconcentrated in vacuo. The product can be purified by standard means,e.g., by chromatography using silica gel (e.g., flash columnchromatography).

Alternatively, N--O compounds can be made from intermediate: ##STR48##by the above oxidation procedure with m-chloroperoxybenzoic acid and##STR49## wherein Q is a protecting group, e.eg., BOC. After oxidationthe protecting group is removed by techniques well known in the art. TheN--O intermediate are then reacted further to produce the compounds ofthe invention.

Compounds of Formula 12.0 include the compound of Formula 12.1:##STR50## The compound of Formula 12.1 is prepared by methods known inthe art, for example by methods disclosed in WO 95/10516, in U.S. Pat.No. 5,151,423 and those described below. The above intermediate compoundcan also be prepared by a procedure comprising the following steps:

(a) reacting an amide of the formula ##STR51## wherein R^(11a) is Br,R^(5a) is hydrogen and R^(6a) is C₁ -C₆ alkyl, aryl or heteroaryl;R^(5a) is C₁ -C₆ alkyl, aryl or heteroaryl and R^(6a) is hydrogen;R^(5a) and R^(6a) are independently selected from the group consistingof C₁ -C₆ alkyl and aryl; or R^(5a) and R^(6a), together with thenitrogen to which they are attached, form a ring comprising 4 to 6carbon atoms or comprising 3 to 5 carbon atoms and one hetero moietyselected from the group consisting of --O-- and --NR^(9a) --, whereinR^(9a) is H, C₁ -C₆ alkyl or phenyl;

with a compound of the formula ##STR52## wherein R^(1a), R^(2a), R^(3a)and R^(4a) are are independently selected from the group consisting ofhydrogen and halo and R^(7a) is Cl or Br, in the presence of a strongbase to obtain a compound of the formula ##STR53## (b) reacting acompound of step (a) with (i) POCl₃ to obtain a cyano compound of theformula ##STR54## (ii) DIBALH to obtain an aldehyde of the formula##STR55## (c) reacting the cyano compound or the aldehyde with apiperidine derivative of the formula ##STR56## wherein L is a leavinggroup selected from the group consisting of Cl and Br, to obtain aketone or an alcohol of the formula below, respectively: ##STR57##(d)(i) cyclizing the ketone with CF₃ SO₃ H to obtain a compound ofFormula 13.0a wherein the dotted line represents a double bond; or

(d)(ii) cyclizing the alcohol with polyphosphoric acid to obtain anIntermediate compound wherein the dotted line represents a single bond.

Methods for preparing the Intermediate compounds disclosed in WO95/10516, U.S. Pat. No. 5,151,423 and described below employ a tricyclicketone intermediate. Such intermediates of the formula ##STR58## whereinR^(11b), R^(1a), R^(2a), R^(3a) and R^(4a) are independently selectedfrom the group consisting of hydrogen and halo, can be prepared by thefollowing process comprising:

(a) reacting a compound of the formula ##STR59## (i) with an amine ofthe formula NHR^(5a) R^(6a), wherein R^(5a) and R^(6a) are as defined inthe process above; in the presence of a palladium catalyst and carbonmonoxide to obtain an amide of the formula: ##STR60## (ii) with analcohol of the formula R^(10a) OH, wherein R^(10a) is C₁ -C₆ lower alkylor C₃ -C₆ cycloalkyl, in the presence of a palladium catalyst and carbonmonoxide to obtain the ester of the formula ##STR61## followed byreacting the ester with an amine of formula NHR^(5a) R^(6a) to obtainthe amide;

(b) reacting the amide with an iodo-substituted benzyl compound of theformula ##STR62## wherein R^(1a), R^(2a), R^(3a), R^(4a) and R^(7a) areas defined above, in the presence of a strong base to obtain a compoundof the formula ##STR63## and (c) cyclizing a compound of step (b) with areagent of the formula R^(8a) MgL, wherein R^(8a) is C₁ -C₈ alkyl, arylor heteroaryl and L is Br or Cl, provided that prior to cyclization,compounds wherein R^(5a) or R^(6a) is hydrogen are reacted with asuitable N-protecting group.

(+)-Isomers of compounds of Formula 12.2 ##STR64## can be prepared withhigh enantioselectivity by using a process comprising enzyme catalyzedtransesterification. Preferably, a racemic compound of Formula 12.3##STR65## is reacted with an enzyme such as Toyobo LIP-300 and anacylating agent such as trifluoroethly isobutyrate; the resultant(+)-amide is then isolated form the (-)-enantomeric amine by techniqueswell known in the art, and then the (+)-amide is hydrolyzed, for exampleby refluxing with an acid such as H₂ SO₄, and the resulting compound isthen reduced with DIBAL by techniques well known in the art to obtainthe corresponding optically enriched (+)-isomer of Formula 12.2.Alternatively, a racemic compound of Formula 12.3, is first reduced tothe corresponding racemic compound of Formula 12.2 and then treated withthe enzyme (Toyobo LIP-300) and acylating agent as described above toobtain the (+)-amide, which is hydrolyzed to obtain the opticallyenriched (+)-isomer.

Those skilled in the art will appreciate that the compounds of formula1.0 having other R¹, R², R³, and R⁴ substituents may be made by theabove enzyme process.

To produce the compounds of Formula 1.0, wherein W is ##STR66## thecompounds of Formulas 14.0 or 15.0 are reacted with the appropriatehaloacetamide or haloacetate: ##STR67## respectively, wherein X is halo(e.g., Br or Cl) to alkylate the amine nitrogen of the piperidine ring.This reaction is conducted according to procedures well known in theart. For example, the compounds of Formulas 14.0 or 15.0 are reactedwith the appropriate haloacetamide in a suitable organic solvent (e.g.,DMF) with a suitable base (e.g., Na₂ CO₃).

For example, reaction of the compound of the formula ##STR68## with thehaloacetamides or haloacetates ##STR69## wherein X is Br or Cl, in DMFwith Na₂ CO₃ yields compounds of the formulas ##STR70## respectively.

Compounds of the invention are exemplified by the following examples,which should not be construed to limit the scope of the disclosure.

PREPARATIVE EXAMPLE 1 ##STR71## Step A: ##STR72##

Combine 10 g (60.5 mmol) of ethyl 4-pyridylacetate and 120 mL of dry CH₂Cl₂ at -20° C., add 10.45 g (60.5 mmol) of MCPBA and stir at -20° C. for1 hour and then at 25° C. for 67 hours. Add an additional 3.48 g (20.2mmoles) of MCPBA and stir at 25° C. for 24 hours. Dilute with CH₂ Cl₂and wash with saturated NaHCO₃ (aqueous) and then water. Dry over MgSO₄,concentrate in vacuo to a residue, and chromatograph (silica gel,2%-5.5% (10% NH₄ OH in MeOH)/CH₂ Cl₂)to give 8.12 g of the productcompound. Mass Spec.: MH⁺ =182.15

Step B: ##STR73##

Combine 3.5 g (19.3 mmol) of the product of Step A, 17.5 mL of EtOH and96.6 mL of 10% NaOH (aqueous) and heat the mixture at 67° C. for 2hours. Add 2 N HCl (aqueous) to adjust to pH=2.37 and concentrate invacuo to a residue. Add 200 mL of dry EtOH, filter through celite® andwash the filter cake with dry EtOH (2×50 ml). Concentrate the combinedfiltrates in vacuo to give 2.43 g of the title compound.

PREPARATIVE EXAMPLE 2 ##STR74##

The title compound is prepared via the process disclosed in PCTInternational Publication No. WO95/10516.

PREPARATIVE EXAMPLE 3 ##STR75## Step A: ##STR76##

Combine 14.95 g (39 mmol) of8-chloro-11-(1-ethoxy-carbonyl-4-piperidinyl)-11H-benzo[5,6]cyclohepta[1,2-b]pyridineand 150 mL of CH₂ Cl₂, then add 13.07 g (42.9 mmol) of (nBu)₄ NNO₃ andcool the mixture to 0° C. Slowly add (dropwise) a solution of 6.09 mL(42.9 mmol) of TFAA in 20 mL of CH₂ Cl₂ over 1.5 hours. Keep the mixtureat 0° C. overnight, then wash successively with saturated NaHCO₃(aqueous), water and brine. Dry the organic solution over Na₂ SO₄,concentrate in vacuo to a residue and chromatograph the residue (silicagel, EtOAc/hexane gradient) to give 4.32 g and 1.90 g of the two productcompounds 3A(i) and 3A(ii), respectively.

Mass Spec. for compound 3A(i): MH⁺ =428.2;

Mass Spec. for compound 3A(ii): MH⁺ =428.3.

Step B: ##STR77##

Combine 22.0 g (51.4 mmol) of the product 3A(i) from Step A, 150 mL of85% EtOH (aqueous), 25.85 g (0.463 mole) of Fe powder and 2.42 g (21.8mmol) of CaCl₂, and heat at reflux overnight. Add 12.4 g (0.222 mole) ofFe powder and 1.2 g (10.8 mmol) of CaCl₂ and heat at reflux for 2 hours.Add another 12.4 g (0.222 mole) of Fe powder and 1.2 g (10.8 mmol) ofCaCl₂ and heat at reflux for 2 hours more. Filter the hot mixturethrough celite®, wash the celite® with 50 mL of hot EtOH and concentratethe filtrate in vacuo to a residue. Add 100 mL of anhydrous EtOH,concentrate to a residue and chromatograph the residue (silica gel,MeOH/CH₂ Cl₂ gradient) to give 16.47 g of the product compound.

Step C: ##STR78##

Combine 16.47 g (41.4 mmol) of the product from Step B, and 150 mL of48% HBr (aqueous) and cool to -3° C. Slowly add (dropwise) 18 mL ofbromine, then slowly add (dropwise) a solution of 8.55 g (0.124 mole) ofNaNO₂ in 85 mL of water. Stir for 45 minutes at -3° to 0° C., thenadjust to pH=10 by adding 50% NaOH (aqueous). Extract with EtOAc, washthe extracts with brine and dry the extracts over Na₂ SO₄. Concentrateto a residue and chromatograph (silica gel, EtOAc/hexane gradient) togive 10.6 g and 3.28 g of the two product compounds 3C(i) and 3C(ii),respectively.

Mass Spec. for compound 3C(i): MH⁺ =461.2;

Mass Spec. for compound 3C(ii): MH⁺ =539.

Step D: ##STR79##

Hydrolyze the product 3C(i) of Step C by dissolving in concentrated HCland heating to about 100° C. for @ 16 hours. Cool the mixture, theneutralize with 1 M NaOH (aqueous). Extract with CH₂ Cl₂, dry theextracts over MgSO₄, filter and concentrate in vacuo to the titlecompound.

Mass Spec.: MH⁺ =466.9.

Step E: ##STR80##

Dissolve 1.160 g (2.98 mmol) of the title compound from Step D in 20 mLof DMF, stir at room temperature, and add 0.3914 g (3.87 mmol) of4-methyl-morpholine, 0.7418 g (3.87 mmol) of DEC, 0.5229 g (3.87 mmol)of HOBT, and 0.8795 g (3.87 mmol) of1-N-t-butoxycarbonyl-piperidinyl-4-acetic acid. Stir the mixture at roomtemperature for 2 days, then concentrate in vacuo to s residue andpartition the residue between CH₂ Cl₂ and water. Wash the organic phasesuccessively with saturated NaHCO₃ (aqueous), 10% NaH₂ PO₄ (aqueous) andbrine. Dry the organic phase over MgSO₄, filter and concentrate in vacuoto a residue. Chromatograph the residue (silica gel, 2% MeOH/CH₂ Cl₂+NH₃) to give 1.72 g of the product. m.p.=94.0-94.5° C., Mass Spec.: MH⁺=616.3,

elemental analysis: calculated--C, 60.54; H, 6.06; N, 6.83 found--C,59.93; H, 6.62; N, 7.45.

Step F: ##STR81##

Combine 1.67 g (2.7 mmol) of the product of Step E and 20 mL of CH₂ Cl₂and stir at 0° C. Add 20 mL of TFA, stir the mixture for 2 hours, thenbasify the mixture with 1 N NaOH (aqueous). Extract with CH₂ Cl₂, drythe organic phase over MgSO₄, filter and concentrate in vacuo to give1.16 g of the product. m.p.=140.2-140.8° C., Mass Spec.: MH⁺ =516.2.

PREPARATIVE EXAMPLE 4 ##STR82## Step A: ##STR83##

Combine 25.86 g (55.9 mmol) of4-(8-chloro-3-bromo-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylicacid ethyl ester and 250 mL of concentrated H₂ SO₄ at -5° C., then add4.8 g (56.4 mmol) of NaNO₃ and stir for 2 hours. Pour the mixture into600 g of ice and basify with concentrated NH₄ OH (aqueous). Filter themixture, wash with 300 mL of water, then extract with 500 mL of CH₂ Cl₂.Wash the extract with 200 mL of water, dry over MgSO₄, then filter andconcentrate in vacuo to a residue. Chromatograph the residue (silicagel, 10% EtOAc/CH₂ Cl₂) to give 24.4 g (86% yield) of the product.m.p.=165-167° C., Mass Spec.: MH⁺ =506 (Cl),

elemental analysis: calculated--C, 52.13; H, 4.17; N, 8.29 found--C,52.18; H, 4.51; N, 8.16.

Step B: ##STR84##

Combine 20 g (40.5 mmol) of the product of Step A and 200 mL ofconcentrated H₂ SO₄ at 20° C., then cool the mixture to 0° C. Add 7.12 g(24.89 mmol) of 1,3-dibromo-5,5-dimethyl-hydantoin to the mixture andstir for 3 hours at 20° C. Cool to 0° C., add an additional 1.0 g (3.5mmol) of the dibromohydantoin and stir at 20° C. for 2 hours. Pour themixture into 400 g of ice, basify with concentrated NH₄ OH (aqueous) at0° C., and collect the resulting solid by filtration. Wash the solidwith 300 mL of water, slurry in 200 mL of acetone and filter to provide19.79 g (85.6% yield) of the product. m.p.=236-237° C., Mass Spec.: MH⁺=584 (Cl),

elemental analysis: calculated--C, 45.1 1; H, 3.44; N, 7.17 found--C,44.95; H, 3.57; N, 7.16.

Step C: ##STR85##

Combine 25 g (447 mmol) of Fe filings, 10 g (90 mmol) of CaCl₂ and asuspension of 20 g (34.19 mmol) of the product of Step B in 700 mL of90:10 EtOH/water at 50° C. Heat the mixture at reflux overnight, filterthrough Celite® and wash the filter cake with 2×200 mL of hot EtOH.Combine the filtrate and washes, and concentrate in vacuo to a residue.Extract the residue with 600 mL of CH₂ Cl₂, wash with 300 mL of waterand dry over MgSO₄. Filter and concentrate in vacuo to a residue, thenchromatograph (silica gel, 30% EtOAc/CH₂ Cl₂) to give 11.4 g (60% yield)of the product. m.p.=211-212° C.,

Mass Spec.: MH⁺ =554 (Cl),

elemental analysis: calculated--C, 47.55; H, 3.99; N, 7.56 found--C,47.45; H, 4.31; N, 7.49.

Step D: ##STR86##

Slowly add (in portions) 20 g (35.9 mmol) of the product of Step C to asolution of 8 g (116 mmol) of NaNO₂ in 120 mL of concentrated HCl(aqueous) at -10° C. Stir the resulting mixture at 0° C. for 2 hours,then slowly add (dropwise) 150 mL (1.44 mole) of 50% H₃ PO₂ at 0° C.over a 1 hour period. Stir at 0° C. for 3 hours, then pour into 600 g ofice and basify with concentrated NH₄ OH (aqueous). Extract with 2×300 mLof CH₂ Cl₂, dry the extracts over MgSO₄, then filter and concentrate invacuo to a residue. Chromatograph the residue (silica gel, 25%EtOAc/hexanes) to give 13.67 g (70% yield) of the product. m.p.=163-165°C., Mass Spec.: MH⁺ =539 (Cl),

elemental analysis: calculated--C, 48.97; H, 4.05; N, 5.22 found--C,48.86; H, 3.91; N, 5.18.

Step E: ##STR87##

Combine 6.8 g (12.59 mmol) of the product of Step D and 100 mL ofconcentrated HCl (aqueous) and stir at 85° C. overnight. Cool themixture, pour it into 300 g of ice and basify with concentrated NH₄ OH(aqueous). Extract with 2×300 mL of CH₂ Cl₂, then dry the extracts overMgSO₄. Filter, concentrate in vacuo to a residue, then chromatograph(silica gel, 10% MeOH/EtOAc+2% NH₄ OH (aqueous)) to give 5.4 g (92%yield) of the title compound. m.p.=172-174° C., Mass Spec.: MH⁺ =467(FAB),

elemental analysis: calculated--C, 48.69; H, 3.65; N, 5.97 found--C,48.83; H, 3.80; N, 5.97.

Step F:

Following essentially the same procedure as Step C of PreparativeExample 5 below, the title compound from Step E above is reacted with1-N-t-butoxycarbonylpiperidinyl-4-acetic acid to produce the compound##STR88## Step G:

Following essentially the same procedure as Step D of PreparativeExample 5 below, the title compound from Step F above is deprotected toyield the title compound of Preparative Example 4.

PREPARATIVE EXAMPLE 5 ##STR89## Step A: ##STR90##

Hydrolyze 2.42 g of4-(8-chloro-3-bromo-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylicacid ethyl ester via substantially the same procedure as described inPreparative Example 3, Step D, to give 1.39 g (69% yield) of theproduct.

Step B: ##STR91##

Combine 1 g (2.48 mmol) of the product of Step A and 25 mL of drytoluene, add 2.5 mL of 1 M DIBAL in toluene and heat the mixture atreflux. After 0.5 hours, add another 2.5 mL of 1 M DIBAL in toluene andheat at reflux for 1 hour. (The reaction is monitored by TLC using 50%MeOH/CH₂ Cl₂ +NH₄ OH (aqueous).) Cool the mixture to room temperature,add 50 mL of 1 N HCl (aqueous) and stir for 5 min. Add 100 mL of 1 NNaOH (aqueous), then extract with EtOAc (3×150 mL). Dry the extractsover MgSO₄, filter and concentrate in vacuo to give 1.1 g of the titlecompound.

Step C: ##STR92##

Combine 0.501 g (1.28 mmol) of the title compound of Step B and 20 mL ofdry DMF, then add 0.405 g (1.664 mmol) of1-N-t-butoxycarbonylpiperidinyl-4-acetic acid, 0.319 g (1.664 mmol) ofDEC, 0.225 g (1.664 mmol) of HOBT, and 0.168 g (1.664 mmol) of4-methylmorpholine and stir the mixture at room temperature overnight.Concentrate the mixture in vacuo to a residue, then partition theresidue between 150 mL of CH₂ Cl₂ and 150 mL of saturated NaHCO₃(aqueous). Extract the aqueous phase with another 150 mL of CH₂ Cl₂. Drythe organic phase over MgSO₄, and concentrate in vacuo to a residue.Chromatograph the residue (silica gel, 500 mL hexane, 1 L of 1% MeOH/CH₂Cl₂ +0.1% NH₄ OH (aqueous), then 1 L of 2% MeOH/CH₂ Cl₂ +0.1% NH₄ OH(aqueous)) to give 0.575 g of the product. m.p.=115°-125° C.; MassSpec.: MH⁺ =616.

Step D: ##STR93## Combine 0.555 g (0.9 mmol) of the product of Step Cand 15 mL of CH₂ Cl₂ and cool the mixture to 0° C. Add 15 mL of TFA andstir at 0° C. for 2 hours. Concentrate in vacuo at 40-45° C. to aresidue, then partition the residue between 150 mL of CH₂ Cl₂ and 100 mLof saturated NaHCO₃ (aqueous). Extract the aqueous layer with 100 mL ofCH₂ Cl₂, combine the extracts and dry over MgSO₄. Concentrate in vacuoto give 0.47 g of the product.

m.p.=140°-150° C.; Mass Spec.: MH⁺ =516.

PREPARATIVE EXAMPLE 6 ##STR94## Step A: ##STR95##

Combine 16.6 g (0.03 mole) of the product of Preparative Example 4, StepD, with a 3:1 solution of CH₃ CN and water (212.65 mL CH₃ CN and 70.8 mLof water) and stir the resulting slurry overnight at room temperature.Add 32.833 g (0.153 mole) of NaIO₄ and then 0.31 g (2.30 mmol) of RuO₂and stir at room temperature give 1.39 g (69% yield) of the product.(The addition of RuO is accompanied by an exothermic reaction and thetemperature climbs from 20° to 30° C.) Stir the mixture for 1.3 hrs.(temperature returned to 25° C. after about 30 min.), then filter toremove the solids and wash the solids with CH₂ Cl₂. Concentrate thefiltrate in vacuo to a residue and dissolve the residue in CH₂ Cl₂.Filter to remove insoluble solids and wash the solids with CH₂ Cl₂. Washthe filtrate with water, concentrate to a volume of about 200 mL andwash with bleach, then with water. Extract with 6 N HCl (aqueous). Coolthe aqueous extract to 0° C. and slowly add 50% NaOH (aqueous) to adjustto pH=4 while keeping the temperature <30° C. Extract twice with CH₂Cl₂, dry over MgSO₄ and concentrate in vacuo to a residue. Slurry theresidue in 20 mL of EtOH and cool to 0° C. Collect the resulting solidsby filtration and dry the solids in vacuo to give 7.95 g of the product.¹ H NMR (CDCl₃, 200 MHz): 8.7 (s, 1H); 7.85 (m, 6H); 7.5 (d, 2H); 3.45(m, 2H); 3.15 (m, 2H).

Step B: ##STR96##

Combine 21.58 g (53.75 mmol) of the product of Step A and 500 mL of ananhydrous 1:1 mixture of EtOH and toluene, add 1.43 g (37.8 mmol) ofNaBH₄ and heat the mixture at reflux for 10 min. Cool the mixture to 0°C., add 100 mL of water, then adjust to pH≈4-5 with 1 M HCl (aqueous)while keeping the temperature <10° C. Add 250 mL of EtOAc and separatethe layers. Wash the organic layer with brine (3×50 mL) then dry overNa₂ SO₄. Concentrate in vacuo to a residue (24.01 g) and chromatographthe residue (silica gel, 30% hexane/CH₂ Cl₂) to give the product. Impurefractions were purified by rechromatography. A total of 18.57 g of theproduct was obtained. ¹ H NMR (DMSO-d₆, 400 MHz): 8.5 (s, 1H); 7.9 (s,1H); 7.5 (d of d, 2H); 6.2 (s, 1H); 6.1 (s, 1H); 3.5 (m, 1H); 3.4 (m,1H); 3.2 (m, 2H).

Step C: ##STR97##

Combine 18.57 g (46.02 mmol) of the product of Step B and 500 mL ofCHCl₃, then add 6.70 mL (91.2 mmol) of SOCl₂, and stir the mixture atroom temperature for 4 hrs. Add a solution of 35.6 g (0.413 mole) ofpiperazine in 800 mL of THF over a period of 5 min. and stir the mixturefor 1 hr. at room temperature. Heat the mixture at reflux ovemight, thencool to room temperature and dilute the mixture with 1 L of CH₂ Cl₂.Wash with water (5×200 mL), and extract the aqueous wash with CHCl₃(3×100 mL). Combine all of the organic solutions, wash with brine (3×200mL) and dry over MgSO₄. Concentrate in vacuo to a residue andchromatograph (silica gel, gradient of 5%, 7.5%, 10% MeOH/CH₂ Cl₂ +NH₄OH) to give 18.49 g of the title compound as a racemic mixture.

Step D--Separation of Enantiomers: ##STR98##

The racemic title compound of Step C is separated by preparative chiralchromatography (Chiralpack AD, 5 cm×50 cm column, flow rate 100 mL/min.,20% iPrOH/hexane+0.2% diethylamine), to give 9.14 g of the (+)-isomerand 9.30 g of the (-)-isomer.

Physical chemical data for (+)-isomer: m.p.=74.5°-77.5° C.; Mass Spec.MH⁺ =471.9; [α]_(D) ²⁵ =+97.4° (8.48 mg/2 mL MeOH).

Physical chemical data for (-)-isomer: m.p.=82.9°-84.5° C.; Mass Spec.MH⁺ =471.8; [α]_(D) ²⁵ =-97.4° (8.32 mg/2 mL MeOH).

Step E: ##STR99##

Combine 3.21 g (6.80 mmol) of the (-)-isomer product of Step D and 150mL of anhydrous DMF. Add 2.15 g (8.8 mmol) of1-N-t-butoxycarbonylpiperidinyl-4-acetic acid, 1.69 g (8.8 mmol) of DEC,1.19 g (8.8 mmol) of HOBT and 0.97 mL (8.8 mmol) of N-methylmorpholineand stir the mixture at room temperature overnight. Concentrate in vacuoto remove the DMF and add 50 mL of saturated NaHCO₃ (aqueous). Extractwith CH₂ Cl₂ (2×250 mL), wash the extracts with 50 mL of brine and dryover MgSO₄. Concentrate in vacuo to a residue and chromatograph (silicagel, 2% MeOH/CH₂ Cl₂ +10% NH₄ OH) to give 4.75 g of the product.m.p.=75.7°-78.5° C.; Mass Spec.: MH⁺ =697; [α]_(D) ²⁵ =-5.5° (6.6 mg/2mL MeOH).

Step F: ##STR100##

Combine 4.70 g (6.74 mmol) of the product of Step E and 30 mL of MeOH,then add 50 mL of 10% H₂ SO₄ /dioxane in 10 mL aliquots over a 1 hr.period. Pour the mixture into 50 mL of water and add 15 mL of 50% NaOH(aqueous) to adjust to pH≈10-11. Filter to remove the resulting solidsand extract the filtrate with CH₂ Cl₂ (2×250 mL). Concentrate theaqueous layer in vacuo to remove the MeOH and extract again with 250 mLof CH₂ Cl₂. Dry the combined extracts over MgSO₄ and concentrate invacuo to give the product. m.p.=128.1°-131.5° C.; Mass Spec.: MH⁺ =597;[α]_(D) ²⁵ =-6.02° (9.3 mg/2 mL MeOH).

PREPARATIVE EXAMPLE 7 ##STR101## Step A: ##STR102##

Combine 15 g (38.5 mmol) of4-(8-chloro-3-bromo-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylicacid ethyl ester and 150 mL of concentrated H₂ SO₄ at -5° C., then add3.89 g (38.5 mmol) of KNO₃ and stir for 4 hours. Pour the mixture into 3L of ice and basify with 50% NaOH (aqueous). Extract with CH₂ Cl₂, dryover MgSO₄, then filter and concentrate in vacuo to a residue.Recrystallize the residue from acetone to give 6.69 g of the product. ¹H NMR (CDCl₃, 200 MHz): 8.5 (s, 1H); 7.75 (s, 1H); 7.6 (s, 1H); 7.35 (s,1H); 4.15 (q, 2H); 3.8 (m, 2H); 3.5-3.1 (m, 4H); 3.0-2.8 (m, 2H);2.6-2.2 (m, 4H); 1.25 (t, 3H).

Step B: ##STR103##

Combine 6.69 g (13.1 mmol) of the product of Step A and 100 mL of 85%EtOH/water, then add 0.66 g (5.9 mmol) of CaCl₂ and 6.56 g (117.9 mmol)of Fe and heat the mixture at reflux overnight. Filter the hot reactionmixture through celite® and rinse the filter cake with hot EtOH.Concentrate the filtrate in vacuo to give 7.72 g of the product. MassSpec.: MH⁺ =478.0.

Step C: ##STR104##

Combine 7.70 g of the product of Step B and 35 mL of HOAc, then add 45mL of a solution of Br₂ in HOAc and stir the mixture at room temperatureovernight. Add 300 mL of 1 N NaOH (aqueous), then 75 mL of 50% NaOH(aqueous) and extract with EtOAc. Dry the extract over MgSO₄ andconcentrate in vacuo to a residue. Chromatograph the residue (silicagel, 20%-30% EtOAc/hexane) to give 3.47 g of the product (along withanother 1.28 g of partially purified product). Mass Spec.: MH⁺ =555.9.

¹ H NMR (CDCl₃, 300 MHz): 8.5 (s, 1H); 7.5 (s, 1H); 7.15 (s, 1H); 4.5(s, 2H); 4.15 (m, 3H); 3.8 (br s, 2H); 3.4-3.1 (m, 4H); 9-2.75 (m, 1H);2.7-2.5 (m, 2H); 2.4-2.2 (m, 2H); 1.25 (m, 3H).

Step D: ##STR105##

Combine 0.557 g (5.4 mmol) of t-butylnitrite and 3 mL of DMF, and heatthe mixture at to 60°-70° C. Slowly add (dropwise) a mixture of 2.00 g(3.6 mmol) of the product of Step C and 4 mL of DMF, then cool themixture to room temperature. Add another 0.64 mL of t-butylnitrite at40° C. and reheat the mixture to 60°-70° C. for 0.5 hrs. Cool to roomtemperature and pour the mixture into 150 mL of water. Extract with CH₂Cl₂, dry the extract over MgSO₄ and concentrate in vacuo to a residue.

Chromatograph the residue (silica gel, 10%-20% EtOAc/hexane) to give0.74 g of the product. Mass Spec.: MH⁺ =541.0.

¹ H NMR (CDCl3, 200 MHz): 8.52 (s, 1H); 7.5 (d, 2H); 7.2 (s, 1H); 4.15(q, 2H); 3.9-3.7 (m, 2H); 3.5-3.1 (m, 4H); 3.0-2.5 (m, 2H); 2.4-2.2 (m,2H); 2.1-1.9 (m, 2H); 1.26 (t, 3H).

Step E: ##STR106##

Combine 0.70 g (1.4 mmol) of the product of Step D and 8 mL ofconcentrated HCl (aqueous) and heat the mixture at reflux overnight. Add30 mL of 1 N NaOH (aqueous), then 5 mL of 50% NaOH (aqueous) and extractwith CH₂ Cl₂. Dry the extract over MgSO₄ and concentrate in vacuo togive 0.59 g of the title compound. Mass Spec.: M⁺ =468.7.m.p.=123.9°-124.2° C.

Step F: ##STR107##

React 6.0 g (12.8 mmol) of the title compound from Step E and with 3.78g (16.6 mmol) of 1-N-t-butoxycarbonylpiperidinyl-4-acetic acid usingsubstantially the same procedures as described for Preparative Example5, Step C, to give 8.52 g of the product. Mass Spec.: MH⁺ =694.0 (FAB).

¹ H NMR (CDCl₃, 200 MHz): 8.5 (d, 1H); 7.5 (d, 2H); 7.2 (d, 1H);4.15-3.9 (m, 3H); 3.8-3.6 (m, 1H); 3.5-3.15 (m, 3H); 2.9 (d, 2H);2.8-2.5 (m, 4H); 2.4-1.8 (m, 6H); 1.8-1.6 (br d, 2H); 1.4 (s, 9H);1.25-1.0 (m, 2H).

Step G: ##STR108##

Combine 8.50 g of the product of Step F and 60 mL of CH₂ Cl₂, then coolto 0° C. and add 55 mL of TFA. Stir the mixture for 3 h at 0° C., thenadd 500 mL of 1 N NaOH (aqueous) followed by 30 mL of 50% NaOH(aqueous). Extract with CH₂ Cl₂, dry over MgSO₄ and concentrate in vacuoto give 7.86 g of the product. Mass Spec.: M⁺ =593.9 (FAB). ¹ H NMR(CDCl₃, 200 MHz): 8.51 (d, 1H); 7.52 (d of d, 2H); 7.20 (d, 1H);4.1-3.95 (m, 2H); 3.8-3.65 (m, 2H); 3.5-3.05 (m, 5H); 3.0-2.5 (m, 6H);2.45-1.6 (m, 6H);1.4-1.1 (m, 2H).

PREPARATIVE EXAMPLE 8 ##STR109## Step A: ##STR110##

Prepare a solution of 8.1 g of the title compound from PreparativeExample 7, Step E, in toluene and add 17.3 mL of a 1M solution of DIBALin toluene. Heat the mixture at reflux and slowly add (dropwise) another21 mL of 1 M DIBAL/toluene solution over a period of 40 min. Cool thereaction mixture to about 0° C. and add 700 mL of 1 M HCl (aqueous).Separate and discard the organic phase. Wash the aqueous phase with CH₂Cl₂, discard the extract, then basify the aqueous phase by adding 50%NaOH (aqueous). Extract with CH₂ Cl₂, dry the extract over MgSO₄ andconcentrate in vacuo to give 7.30 g of the title compound, which is aracemic mixture of enantiomers.

Step B--Separation of Enantiomers: ##STR111##

The racemic title compound of Step A is separated by preparative chiralchromatography (Chiralpack AD, 5 cm×50 cm column, using 20%iPrOH/hexane+0.2% diethylamine), to give the (+)-isomer and the(-)-isomer of the title compound.

Physical chemical data for (+)-isomer: m.p.=148.8° C.; Mass Spec. MH⁺=469; [α]_(D) ²⁵ =+65.60 (12.93 mg/2 mL MeOH).

Physical chemical data for (-)-isomer: m.p.=112° C.; Mass Spec. MH⁺=469; [α]_(D) ²⁵ =-65.2° (3.65 mg/2 mL MeOH).

Step C: ##STR112##

React 1.33 g of the (+)-isomer of the title compound of PreparativeExample 8, Step B, with 1.37 g of1-N-t-butoxy-carbonylpiperidinyl-4-acetic acid using substantially thesame procedures as described for Preparative Example 5, Step C, to give2.78 g of the product. Mass Spec.: MH⁺ =694.0 (FAB); [α]_(D) ²⁵ =+34.1°(5.45 mg/2 mL, MeOH).

Step D: ##STR113##

Treat 2.78 g of the product of Step C via substantially the sameprocedure as described for Preparative Example 5, Step D, to give 1.72 gof the product. m.p.=104.1° C.; Mass Spec.: MH⁺ =594; [α]_(D) ²⁵ =+53.4°(11.42 mg/2 mL, MeOH).

PREPARATIVE EXAMPLE 9 ##STR114## Step A: ##STR115##

Combine 40.0 g (0.124 mole) of the starting ketone and 200 mL of H₂ SO₄and cool to 0° C. Slowly add 13.78 g (0.136 mole) of KNO₃ over a periodof 1.5 hrs., then warm to room temperature and stir overnight. Work upthe reaction using substantially the same procedure as described forPreparative Example 4, Step A. Chromatograph (silica gel, 20%, 30%, 40%,50% EtOAc/hexane, then 100% EtOAc) to give 28 g of the 9-nitro product,along with a smaller quantity of the 7-nitro product and 19 g of amixture of the 7-nitro and 9-nitro compounds.

Step B: ##STR116##

React 28 g (76.2 mmol) of the 9-nitro product of Step A, 400 mL of 85%EtOH/water, 3.8 g (34.3 mmol) of CaCl₂ and 38.28 g (0.685 mole) of Feusing substantially the same procedure as described for PreparativeExample 4, Step C, to give 24 g of the product

Step C: ##STR117##

Combine 13 9 (38.5 mmol) of the product of Step B, 140 mL of HOAc andslowly add a solution of 2.95 mL (57.8 mmol) of Br₂ in 10 mL of HOAcover a period of 20 min. Stir the reaction mixture at room temperature,then concentrate in vacuo to a residue. Add CH₂ Cl₂ and water, thenadjust to pH=8-9 with 50% NaOH (aqueous). Wash the organic phase withwater, then brine and dry over Na₂ SO₄. Concentrate in vacuo to give11.3 g of the product.

Step D: ##STR118##

Cool 100 mL of concentrated HCl (aqueous) to 0° C., then add 5.61 g(81.4 mmol) of NaNO₂ and stir for 10 min. Slowly add (in portions) 11.3g (27.1 mmol) of the product of Step C and stir the mixture at 0°-3° C.for 2.25 hrs. Slowly add (dropwise) 180 mL of 50% H₃ PO₂ (aqueous) andallow the mixture to stand at 0° C. overnight. Slowly add (dropwise) 150mL of 50% NaOH over 30 min., to adjust to pH=9, then extract with CH₂Cl₂. Wash the extract with water, then brine and dry over Na₂ SO₄.Concentrate in vacuo to a residue and chromatograph (silica gel, 2%EtOAc/CH₂ Cl₂) to give 8.6 g of the product.

Step E: ##STR119##

Combine 8.6 g (21.4 mmol) of the product of Step D and 300 mL of MeOHand cool to 0°-2° C. Add 1.21 g (32.1 mmol) of NaBH₄ and stir themixture at ˜0° C. for 1 hr. Add another 0.121 g (3.21 mmol) of NaBH₄,stir for 2 hr. at 0° C., then let stand overnight at 0° C. Concentratein vacuo to a residue then partition the residue between CH₂ Cl₂ andwater. Separate the organic phase and concentrate in vacuo (50° C.) togive 8.2 g of the product.

Step F: ##STR120##

Combine 8.2 g (20.3 mmol) of the product of Step E and 160 mL of CH₂Cl₂, cool to 0° C., then slowly add (dropwise) 14.8 mL (203 mmol) ofSOCl₂ over a 30 min. period. Warm the mixture to room temperature andstir for 4.5 hrs., then concentrate in vacuo to a residue, add CH₂ Cl₂and wash with 1 N NaOH (aqueous) then brine and dry over Na₂ SO₄.Concentrate in vacuo to a residue, then add dry THF and 8.7 g (101 mmol)of piperazine and stir at room temperature overnight. Concentrate invacuo to a residue, add CH₂ Cl₂, and wash with 0.25 N NaOH (aqueous),water, then brine. Dry over Na₂ SO₄ and concentrate in vacuo to give9.46 g of the crude product. Chromatograph (silica gel, 5% MeOH/CH₂ Cl₂+NH₃) to give 3.59 g of the title compound, as a racemate. ¹ H NMR(CDCl₃, 200 MHz): 8.43 (d, 1H); 7.55 (d, 1H); 7.45 (d, 1H); 7.11 (d,1H); 5.31 (s, 1H); 4.86-4.65 (m, 1H); 3.57-3.40 (m, 1H); 2.98-2.55 (m,6H); 2.45-2.20 (m, 5H).

Step G--Separation of Enantiomers: ##STR121##

The racemic title compound from Step F (5.7 g) is chromatographed asdescribed for Preparative Example 6, Step D, using 30% iPrOH/hexane+0.2%diethylamine, to give 2.88 g of the R-(+)-isomer and 2.77 g of theS-(-)-isomer of the title compound.

Physical chemical data for the R-(+)-isomer: Mass Spec. MH⁺ =470.0;[α]_(D) ²⁵ =+12.1° (10.9 mg/2 mL MeOH).

Physical chemical data for the S-(-)-isomer: Mass Spec. MH⁺ =470.0;[α]_(D) ²⁵ =-13.2° (11.51 mg/2 mL MeOH).

Step H:

Following essentially the same procedure as Preparative Example 5, StepsC and D, the racemic title compound of Preparative Example 9 is obtainedfrom the racemic compound of Step F. Similarly, using the (-)- or(+)-isomer from Step G, the (-)- or (+)-isomer of the title compound ofPreparative Example 9 is obtained, respectively.

PREPARATIVE EXAMPLE 10 ##STR122## Step A: ##STR123##

Combine 13 g (33.3 mmol) of the title compound from Preparative Example4, Step E, and 300 mL of toluene at 20° C., then add 32.5 mL (32.5 mmol)of a 1 M solution of DIBAL in toluene. Heat the mixture at reflux for 1hr., cool to 20° C., add another 32.5 mL of 1 M DIBAL solution and heatat reflux for 1 hr. Cool the mixture to 20° C. and pour it into amixture of 400 g of ice, 500 mL of EtOAc and 300 mL of 10% NaOH(aqueous). Extract the aqueous layer with CH₂ Cl₂ (3×200 mL), dry theorganic layers over MgSO₄, then concentrate in vacuo to a residue.Chromatograph (silica gel, 12% MeOH/CH₂ Cl₂ +4% NH₄ OH) to give 10.4 gof the title compound as a racemate. Mass Spec.: MH⁺ =469 (FAB). partial¹ H NMR (CDCl₃, 400 MHz): 8.38 (s, 1H); 7.57 (s, 1H); 7.27 (d, 1H); 7.06(d, 1H); 3.95 (d, 1H).

Step B--Separation of Enantiomers: ##STR124##

The racemic title compound of Step A is separated by preparative chiralchromatography (Chiralpack AD, 5 cm×50 cm column, using 5%iPrOH/hexane+0.2% diethylamine), to give the (+)-isomer and the(-)-isomer of the title compound.

Physical chemical data for (+)-isomer: Mass Spec. MH⁺ =469 (FAB);[α]_(D) ²⁵ =+43.5° (c=0.402, EtOH); partial ¹ H NMR (CDCl₃, 400 MHz):8.38 (s, 1H); 7.57 (s, 1H); 7.27 (d, 1H); 7.05 (d, 1H); 3.95 (d, 1H).

Physical chemical data for (-)-isomer Mass Spec. MH⁺ =469 (FAB); [α]_(D)²⁵ =41.8° (c=0.328 EtOH); partial ¹ H NMR (CDCl₃, 400 MHz): 8.38 (s,1H); 7.57 (s, 1H); 7.27 (d, 1H); 7.05 (d, 1H); 3.95 (d, 1H).

Step C:

Following the procedure of Preparative Example 9, Step H, the racemiccompound, the (+)-isomer or the (-)-isomer of the title compound ofPreparative Example 10 can be obtained.

PREPARATIVE EXAMPLE 11 ##STR125##

The compound ##STR126## is prepared according to the procedures ofPreparative Example 40 of WO 95/10516 (published Apr. 20, 1995), byfollowing the procedures described in Example 193 of WO 95/10516.

The (+)- and (-)-isomers can be separated by following essentially thesame procedure as Step D of Preparative Example 6.

Physical chemical data for the R-(+)-isomer: ¹³ C NMR (CDCl₃): 155.8(C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2 (C); 135.3 (C); 133.4 (C);132.0 (CH); 129.9 (CH); 125.6 (CH); 119.3 (C); 79.1 (CH); 52.3 (CH₂);52.3 (CH); 45.6 (CH₂); 45.6 (CH₂); 30.0 (CH₂); 29.8 (CH₂). [α]_(D) ²⁵=+25.8° (8.46 mg/2 mL MeOH).

Physical chemical data for the S-(-)-isomer: ¹³ C NMR (CDCl₃): 155.9(C); 146.4 (CH); 140.5 (CH); 140.2 (C); 136.2 (C); 135.3 (C); 133.3 (C);132.0 (CH); 129.9 (CH); 125.5 (CH); 119.2 (C); 79.1 (CH); 52.5 (CH₂);52.5 (CH); 45.7 (CH₂); 45.7 (CH₂); 30.0 (CH₂); 29.8 (CH₂). [α]_(D) ²⁵=-27.9° (8.90 mg/2 mL MeOH).

Following essentially the same procedure as Preparative Example 5, StepsC and D, the racemic compound, (+)-isomer or (-)-isomer of the titlecompound of Preparative Example 11 can be obtained from thecorresponding racemic compound, (+)-isomer or (-)-isomer of the compound##STR127##

EXAMPLE 1 ##STR128##

The compound of Formula 16.0 ##STR129## (Preparative Example 8) (0.11 g,0.19 mmol), anhydrous dimethylformamide (2 mL), 2-bromoacetamide (0.027g, 0.2 mmol) and anhydrous sodium carbonate (0.04 g, 0.38 mmol) werestirred at room temperature overnight. The mixture was diluted withwater, filtered and the solids washed with water. The solids werediluted with dichloromethane, dried over anhydrous magnesium sulfate,filtered and concentrated in vacuo to afford the compound of Formula 2.0as a solid (0.084 g, 68%, mp 131° C.).

EXAMPLE 2 ##STR130## Step A: ##STR131##

To a stirred solution of morpholine (1.62 mL, 18.5 mmol) andtriethylamine (2.62 mL, 18.8 mmol) in anhydrous diethyl ether (100 mL)was added chloroacetyl chloride (1.5 mL, 1.02 eq) dissolved in diethylether (10 mL) at 0° C. After stirring for 0.5 hours, water and 1 Mhydrochloric acid were added and the mixture was shaken. The organicphase was separated and washed with brine and 1 N aqueous sodiumhydroxide, then dried over anhydrous magnesium sulfate. Filtration andconcentration in vacuo afforded the title compound (0.05 g) as a lightoil.

Step B: ##STR132##

A mixture of the compound of Formula 16.0 (Preparative Example 8) (0.10g, 0.17 mmol), anhydrous dimethylformamide (4 mL), the title compoundfrom Step A of Example 2 (0.051 g, 0.31 mmol) and anhydrous sodiumcarbonate (0.036 g, 0.34 mmol) were stirred at room temperatureovernight. The mixture was diluted with water, filtered and the solidswashed with water. The solids were diluted with dichloromethane, driedover anhydrous magnesium sulfate, filtered and concentrated in vacuo toafford a solid (0.077 g). Purification by preparative platechromatography (silica gel) using 5% methanol-dichloromethane andconcentrated ammonium hydroxide using 5% methanol-dichloromethane andconcentrated ammonium hydroxide provided the compound of Formula 7.0(0.063 g, 52%, mp 126.9-131.9° C.).

EXAMPLE 3 ##STR133## Step A: ##STR134##

To a stirred solution of t-butyl-1-piperazine carboxylate (2.12 g, 11.4mmol) and triethylamine (1.62 mL, 11.6 mmol) in anhydrous diethyl ether(100 mL) was added chloroacetyl chloride (0.92 mL, 1.02 eq) dissolved indiethyl ether (10 mL) at 0° C. After stirring for 0.5 hours, water and 1M hydrochloric acid were added and the mixture was shaken. The organicphase was separated and washed with brine and 1 N aqueous sodiumhydroxide, then dried over anhydrous magnesium sulfate. Filtration andconcentration in vacuo afforded the title compound (2.37 g) as a lightoil.

Step B: ##STR135##

A mixture of the compound of Formula 16.0 (Preparative Example 8) (0.25g, 0.41 mmol), anhydrous dimethylformamide (5 mL), the title compoundfrom Step A of Example 3 (0.12 g, 0.46 mmol) and anhydrous sodiumcarbonate (0.053 g, 0.50 mmol) were stirred at room temperatureovernight. The mixture was diluted with water, filtered and the solidswashed with water. The solids were diluted with dichloromethane, driedover anhydrous magnesium sulfate, filtered and concentrated in vacuo toafford a solid (0.296 g). Purification by preparative platechromatography (silica gel) using 5% methanol-dichloromethane andconcentrated ammonium hydroxide using 5% methanol-dichloromethane andconcentrated ammonium hydroxide provided the compound of Formula 8.0(0.17 g, 49%, mp 139.1-141.2° C.).

EXAMPLE 4 ##STR136##

To the compound of Formula 8.0 (Example 3) (0.12 g) dissolved inanhydrous dichloromethane (25 mL) was added trifluoroacetic acid (1 mL)and the resulting solution was stirred at room temperature for 2 hour.50% Aqueous sodium hydroxide was added slowly followed bydichloromethane and brine. The mixture was shaken well, the organicphase was washed with water, separated and dried over anhydrousmagnesium sulfate. Filtration and concentration in vacuo provided aresidue which was purified by preparative plate chromatography (silicagel) using 5% methanol-dichloromethane and concentrated ammoniumhydroxide using 5% methanol-dichloromethane and concentrated ammoniumhydroxide affording the compound of Formula 9.0 (0.07 g, 71%, mp130.3-135.2° C.).

EXAMPLE 5 ##STR137##

A mixture of the compound of Formula 16.0 (Preparative Example 8) (0.51g, 0.85 mmol), anhydrous dimethylformamide (10 mL),tertbutylbromoacetate (0.13 mL, 0.89 mmol) and anhydrous sodiumcarbonate (0.18 g, 1.7 mmol) were stirred at room temperature overnight.The mixture was diluted with water, filtered and the solids washed withwater. The solids were diluted with dichloromethane, dried overanhydrous magnesium sulfate, filtered and concentrated in vacuo toafford the compound of Formula 11.0 as a solid (0.47 g, 79%, mp 107-112°C.).

EXAMPLE 6 ##STR138##

Following procedures similar to those of Example 1, the compound ofFormula 16.0 was reacted with 2-chloro-N-methoxy-N-methylacetamide toproduce a compound of Formula 3.0. (Yield 40%, mp 116-120° C.).

EXAMPLE 7 ##STR139##

Following procedures similar to those of Example 1, the compound ofFormula 16.0 was reacted with 2-chloro-N,N-diethylacetamide to produce acompound of Formula 4.0 (Yield 62%, mp 110-114° C.).

EXAMPLE 8 ##STR140##

Following procedures similar to those of Example 1, the compound ofFormula 16.0 was reacted with N-chloroacetyl-benzylamine (i.e.,##STR141## N-chloroacetyl-N-benzylamine) to produce a compound ofFormula 5.0 (Yield 39%, mp 112-116° C.).

EXAMPLE 9 ##STR142##

Following procedures similar to those of Example 1, the compound ofFormula 16.0 was reacted with1-bromoacetamido-1-deoxy-β-D-galactopyranose to produce a compound ofFormula 6.0 (Yield 23%, mp 187.0-189.9° C.).

EXAMPLE 10 ##STR143##

Following procedures similar to those of Example 2, the compound ofFormula 16.0 was reacted with 1-(2-chloroacetyl)-indoline to produce acompound of Formula 10.0.

EXAMPLE 11 ##STR144##

A mixture of the compound of Formula 11.0 (Example 5) (0.40 g),dichloromethane (10 mL) and trifluoroacetic acid (1 mL) was stirred atroom temperature for 10 days. The mixture was neutralized to pH 7 with1N NaOH (aq), diluted with methanol and concentrated in vacuo. thesolids were washed with absolute ethanol, filtered and the filtrate wasconcentrated in vacuo. Dichloromethane saturated with hydrogen chloride(gas) was added to the resulting cream colored foam and after stirringfor 30 minutes at room temperature, the mixture was filtered and thesolid dried under vacuum to provide the compound of Formula 11.1 (1.0 g,MH⁺ =652) along with sodium chloride as an impurity.

EXAMPLE 12 ##STR145##

Dissolve 1.0 equivalent of the (+) product of Preparative Example 8,Step D, in dichloromethane containing 1.1 equivalents of glycidol andstir for 48 hr. Concentrate under vacuum and chromatograph the residueon silica gel to obtain the produce as a white solid.

Assays

FPT IC₅₀ (inhibition of farnesyl protein transferase, in vitro enzymeassay) and COS Cell IC₅₀ (Cell-Based Assay) were determined followingthe assay procedures described in WO 95110516, published Apr. 20, 1995.GGPT IC₅₀ (inhibition of geranylgeranyl protein transferase, in vitroenzyme assay), Cell Mat Assay, and anti-tumor activity (in vivoanti-tumor studies) could be determined by the assay proceduresdescribed in WO 95/10516. The disclosure of WO 95/10516 is incorporatedherein by reference thereto.

Additional assays can be carried out by following essentially the sameprocedure as described above, but with substitution of alternativeindicator tumor cell lines in place of the T24-BAG cells. The assays canbe conducted using either DLD-1-BAG human colon carcinoma cellsexpressing an activated K-ras gene or SW620-BAG human colon carcinomacells expressing an activated K-ras gene. Using other tumor cell linesknown in the art, the activity of the compounds of this inventionagainst other types of cancer cells could be demonstrated.

Soft Agar Assay:

Anchorage-independent growth is a characteristic of tumorigenic celllines. Human tumor cells are suspended in growth medium containing 0.3%agarose and an indicated concentration of a farnesyl transferaseinhibitor. The solution is overlayed onto growth medium solidified with0.6% agarose containing the same concentration of farnesyl transferaseinhibitor as the top layer. After the top layer is solidified, platesare incubated for 10-16 days at 37° C. under 5% CO₂ to allow colonyoutgrowth. After incubation, the colonies are stained by overlaying theagar with a solution of MTT(3-[4,5-dimethyl-thiazol-2-yl]-2,5-diphenyltetrazolium bromide,Thiazolyl blue) (1 mg/mL in PBS). Colonies can be counted and the IC₅₀'s can be determined.

The results are given in Table 1. In Table 1 "Ex. No." stands for"Example Number" and "nM" stands for

                  TABLE 1                                                         ______________________________________                                                           FPT IC.sub.50 (nM)                                         Ex No.   Formula No.                                                                             (H-ras)      COS Cell IC.sub.50 (nM)                       ______________________________________                                        1        2.0       1.8           6                                            2        7.0       5            25                                            3        8.0       18           --                                            4        9.0       4.3          --                                            5        11.0      28           --                                            6        3.0       3.9          20                                            7        4.0       4.3          40                                            8        5.0       16           --                                            9        6.0       3.4          --                                            12       20.0      2.0          <10                                           ______________________________________                                    

Compound 10.0 had an FPT IC₅₀ (H-ras) of 5.4 nM.

The following compounds had the following FPT IC₅₀ (K-ras) results: 2.0,8.6 nM; 3.0, 26 nM; 4.0, 46.0 nM; 5.0, 51.9 nM; 6.0, 17 nM; 7.0, 20 nM;8.0, 42 nM; and 9.0, 17 nM.

The following compounds had the following Cos Cell IC₅₀ results: 5.0,840 nM; 6.0, 300 nM; 8.0, 22 nM; 9.0, 120 nM; and 10.0, 100 nM.

The following compounds had the following Soft Agar FPT IC₅₀ results:2.0, 70 nM; 3.0, 175 nM; 4.0, 300 nM; 6.0, >500 M; 7.0, 100 nM;8.0, >500 nM; 9.0, 500 nM; and 20.0, 60 nM.

For preparing pharmaceutical compositions from the compounds describedby this invention, inert, pharmaceutically acceptable carriers can beeither solid or liquid. Solid form preparations include powders,tablets, dispersible granules, capsules, cachets and suppositories. Thepowders and tablets may be comprised of from about 5 to about 70 percentactive ingredient. Suitable solid carriers are known in the art, e.g.magnesium carbonate, magnesium stearate, talc, sugar, lactose. Tablets,powders, cachets and capsules can be used as solid dosage forms suitablefor oral administration.

For preparing suppositories, a low melting wax such as a mixture offatty acid glycerides or cocoa butter is first melted, and the activeingredient is dispersed homogeneously therein as by stirring. The moltenhomogeneous mixture is then poured into convenient sized molds, allowedto cool and thereby solidify.

Liquid form preparations include solutions, suspensions and emulsions.As an example may be mentioned water or water-propylene glycol solutionsfor parenteral injection.

Liquid form preparations may also include solutions for intranasaladministration.

Aerosol preparations suitable for inhalation may include solutions andsolids in powder form, which may be in combination with apharmaceutically acceptable carrier, such as an inert compressed gas.

Also included are solid form preparations which are intended to beconverted, shortly before use, to liquid form preparations for eitheroral or parenteral administration. Such liquid forms include solutions,suspensions and emulsions.

The compounds of the invention may also be deliverable transdermally.The transdermal compositions can take the form of creams, lotions,aerosols and/or emulsions and can be included in a transdermal patch ofthe matrix or reservoir type as are conventional in the art for thispurpose.

Preferably the compound is administered orally.

Preferably, the pharmaceutical preparation is in unit dosage form. Insuch form, the preparation is subdivided into unit doses containingappropriate quantities of the active component, e.g., an effectiveamount to achieve the desired purpose.

The quantity of active compound in a unit dose of preparation may bevaried or adjusted from about 0.1 mg to 1000 mg, more preferably fromabout 1 mg. to 300 mg, according to the particular application.

The actual dosage employed may be varied depending upon the requirementsof the patient and the severity of the condition being treated.Determination of the proper dosage for a particular situation is withinthe skill of the art. Generally, treatment is initiated with smallerdosages which are less than the optimum dose of the compound.Thereafter, the dosage is increased by small increments until theoptimum effect under the circumstances is reached. For convenience, thetotal daily dosage may be divided and administered in portions duringthe day if desired.

The amount and frequency of administration of the compounds of theinvention and the pharmaceutically acceptable salts thereof will beregulated according to the judgment of the attending clinicianconsidering such factors as age, condition and size of the patient aswell as severity of the symptoms being treated. A typical recommendeddosage regimen is oral administration of from 10 mg to 2000 mg/daypreferably 10 to 1000 mg/day, in two to four divided doses to blocktumor growth. The compounds are non-toxic when administered within thisdosage range.

The following are examples of pharmaceutical dosage forms which containa compound of the invention. The scope of the invention in itspharmaceutical composition aspect is not to be limited by the examplesprovided.

Pharmaceutical Dosage Form Examples

                  EXAMPLE A                                                       ______________________________________                                        Tablets                                                                       No.     Ingredients      mg/tablet                                                                              mg/tablet                                   ______________________________________                                        1.      Active compound  100      500                                         2.      Lactose USP      122      113                                         3.      Corn Starch, Food Grade,                                                                       30       40                                                  as a 10% paste in                                                             Purified Water                                                        4.      Corn Starch, Food Grade                                                                        45       40                                          5.      Magnesium Stearate                                                                             3        7                                                   Total            300      700                                         ______________________________________                                    

Method of Manufacture

Mix Item Nos. 1 and 2 in a suitable mixer for 10-15 minutes. Granulatethe mixture with Item No. 3. Mill the damp granules through a coarsescreen (e.g., 1/4", 0.63 cm) if necessary. Dry the damp granules.

Screen the dried granules if necessary and mix with Item No. 4 and mixfor 10-15 minutes. Add Item No. 5 and mix for 1-3 minutes. Compress themixture to appropriate size and weigh on a suitable tablet machine.

                  EXAMPLE B                                                       ______________________________________                                        Capsules                                                                      No.   Ingredient       mg/capsule mg/capsule                                  ______________________________________                                        1.    Active compound  100        500                                         2.    Lactose USP      106        123                                         3.    Corn Starch, Food Grade                                                                        40         70                                          4.    Magnesium Stearate NF                                                                          7          7                                                 Total            253        700                                         ______________________________________                                    

Method of Manufacture

Mix Item Nos. 1, 2 and 3 in a suitable blender for 10-15 minutes. AddItem No. 4 and mix for 1-3 minutes. Fill the mixture into suitabletwo-piece hard gelatin capsules on a suitable encapsulating machine.

While the present invention has been described in conjunction with thespecific embodiments set forth above, many alternatives, modificationsand variations thereof will be apparent to those of ordinary skill inthe art. All such alternatives, modifications and variations areintended to fall within the spirit and scope of the present invention.

What is claimed is:
 1. A compound of the formula: ##STR146## or apharmaceutically acceptable salt or solvate thereof, wherein: one of a,b, c and d represents N or NR⁹ wherein R⁹ is O⁻, --CH3 or --(CH₂)_(n)CO₂ H wherein n is 1 to 3, and the remaining a, b, c and d groupsrepresent CR¹ or CR² ;each R¹ and each R² is independently selected fromH, halo, --CF₃, --OR¹⁰, --COR¹⁰, --SR¹⁰, --S(O)_(t) R¹¹ (wherein t is 0,1 or 2), --SCN, --N(R¹⁰)₂, --NR¹⁰ R¹¹, --NO₂, --OC(O)R¹⁰, --CO₂ R¹⁰,--OCO₂ R¹¹, --CN, --NHC(O)R¹⁰, --NHSO₂ R¹⁰, --CONHR¹⁰, --CONHCH₂ CH₂ OH,--NR¹⁰ COOR¹¹, ##STR147## --SR¹¹ C(O)OR¹¹, --SR¹¹ N(R⁷⁵)₂ wherein eachR⁷⁵ is independently selected from H and --C(O)OR¹¹,benzotriazol-1-yloxy, tetrazol-5-ylthio, or substitutedtetrazol-5-ylthio, alkynyl, alkenyl or alkyl, said alkyl or alkenylgroup optionally being substituted with halo, --OR¹⁰ or --CO₂ R¹⁰ ; R³and R⁴ are the same or different and each independently represents H,any of the substituents of R¹ and R², or R³ and R⁴ taken togetherrepresent a saturated or unsaturated C₅ -C₇ fused ring to the benzenering (Ring III); R⁵, R⁶, R⁷ and R⁸ each independently represents H,--CF₃, --COR¹⁰, alkyl or aryl, said alkyl or aryl optionally beingsubstituted with --OR¹⁰, --SR¹⁰, --S(O)_(t) R¹¹, --NR¹⁰ COOR¹¹,--N(R¹⁰)₂, --NO₂, --COR¹⁰, --OCOR¹⁰, --OCO₂ R¹¹, --CO₂ R¹⁰, OPO₃ R¹⁰ orR⁵ is combined with R⁶ to represent ═O or ═S and/or R⁷ is combined withR⁸ to represent ═O or ═S; R¹⁰ represents H, alkyl, aryl, or aralkyl; R¹¹represents alkyl or aryl; X represents CH or C, which C contain a doublebond (represented by the dotted line) to carbon atom 11; the dotted linebetween carbon atoms 5 and 6 represents an optional double bond, suchthat when a double bond is present, A and B independently represent--R¹⁰, halo, --OR¹¹, --OCO₂ R¹¹ or --OC(O)R¹⁰, and when no double bondis present between carbon atoms 5 and 6, A and B each independentlyrepresent H₂, --(OR¹¹ )₂ ; H and halo, dihalo, alkyl and H, (alkyl)₂,--H and --OC(O)R¹⁰, H and --OR¹⁰, ═O, aryl and H, ═NOR¹⁰ or--O--(CH₂)_(p) --O-- wherein p is 2, 3 or 4; and W represents a groupselected from: ##STR148## wherein: R¹² is selected from the groupconsisting of: (1) H; (2) alkyl; (3) aryl; (4) arylalkyl; R¹³ isselected from the group consisting of: (1) H; (2) alkyl; (3) alkoxy; (4)heterocycloalkyl; (5) aryl; and (5) aralkyl; R¹⁴ is selected from thegroup consisting of: (1) H; (2) alkyl; (3) aryl; and (4) heteroaryl;ring ##STR149## represents a heterocycloalkyl ring wherein Y representsthe remainder of the ring, said remainder comprising carbon atoms andoptionally a hetero atom selected from the group consisting of: NH,NR¹⁵, O and S, and said remainder optionally having an aryl ring fusedthereto; R¹⁵ represents --C(O)OR¹⁶ ; and R¹⁶ represents alkyl.
 2. Thecompound of claim 1 wherein R² is H; R¹ is selected from the groupconsisting of: Br and Cl; R³ is selected from the group consisting of:Br and Cl; R⁴ is selected from the group consisting of: H, Br and Cl;R⁵, R⁶, R⁷ and R⁸ are H; A and B are each H₂ ; and the optional bondbetween C5 and C6 is absent.
 3. The compound of claim 2 wherein R⁴ is H.4. The compound of claim 2 wherein R⁴ is selected from the groupconsisting of: Cl or Br.
 5. The compound of claim 4 wherein X is CH. 6.The compound of claim 4 wherein W is selected from the group consistingof:(A) ##STR150## wherein: (1) R¹² is selected from the group consistingof: H, alkyl and aralkyl; and (2) R¹³ is selected from the groupconsisting of: H, alkyl, alkoxy, aralkyl, and heterocycloalkyl; (B)##STR151## wherein the heterocycloalkyl ring ##STR152## is selected fromthe group consisting of: ##STR153## and (C) ##STR154## wherein R¹⁴ is Hor alkyl.
 7. The compound of claim 6 wherein X is CH.
 8. The compound ofclaim 7 wherein R¹² is selected from: H, methyl, ethyl or benzyl; R¹³ isselected from: H, methyl, ethyl, methoxy, benzyl, or ##STR155## and R¹⁴represents H or --C(CH₃).
 9. The compound of claim 1 selected from:##STR156## wherein R¹, R³ and R⁴ are each independently selected fromhalo; and A, B, X and W are as defined in claim
 1. 10. The compound ofclaim 9 wherein R¹ is selected from the group consisting of: Br and Cl;R³ and R⁴ are independently selected from the group consiting of: Br andCl; A and B are each H₂ ; and the optional bond between C5 and C6 isabsent.
 11. The compound of claim 10 wherein R¹ is Br; and R³ is Cl; andR⁴ is Br.
 12. The compound of claim 11 wherein X is CH.
 13. The compoundof claim 11 wherein W is selected from the group consisting of:(A)##STR157## wherein: (1) R¹² is selected from the group consisting of: H,alkyl and aralkyl; and (2) R13 is selected from the group consisting of:H, alkyl, alkoxy, aralkyl, and heterocycloalkyl; (B) ##STR158## whereinthe heterocycloalkyl ring ##STR159## is selected from the groupconsisting of: ##STR160## and (C) ##STR161## wherein R¹⁴ is H or alkyl.14. The compound of claim 13 wherein X is CH.
 15. The compound of claim14 wherein R¹² is selected from: H, methyl, ethyl or benzyl; R¹³ isselected from: H, methyl, ethyl, methoxy, benzyl, or ##STR162## and R¹⁴represents H or --C(CH₃).
 16. The compound of claim 14 wherein saidcompound is a compound of the formula: ##STR163## .
 17. A compoundselected from the group consisting of: ##STR164## .
 18. A compoundselected from the group consisting of: ##STR165## .
 19. A compoundselected from the group consisting of: ##STR166## .
 20. A method oftreating tumor cells in a human by inhibition of farnesyl proteintransferase wherein the cells are pancreatic tumor cells, lung cancercells, myeloid leukemia tumor cells, thyroid follicular tumor cells,myelodysplastic tumor cells, epidermal carcinoma tumor cells, bladdercarcinoma tumor cells, colon tumors cells, breast tumor cells orprostate tumor cells comprising administering to the human in needthereof a farnesyl protein transferase inhibiting amount of a compoundof claim
 1. 21. A method of inhibiting farnesyl protein transferase in ahuman comprising the administration to the human in need thereof afarnesyl protein transferase inhibiting amount of the compound ofclaim
 1. 22. A pharmaceutical composition comprising an effective amountof compound of claim 1 in combination with a pharmaceutically acceptablecarrier.
 23. A method of treating tumor cells in a human by inhibitionof farnesyl protein transferase wherein the cells are pancreatic tumorcells, lung cancer cells, myeloid leukemia tumor cells, thyroidfollicular tumor cells, myelodysplastic tumor cells, epidermal carcinomatumor cells, bladder carcinoma tumor cells, colon tumors cells, breasttumor cells or prostate tumor cells comprising administering to thehuman in need thereof a farnesyl protein transferase inhibitingeffective amount of a compound of claim
 17. 24. A method of inhibitingfarnesyl protein transferase in a human comprising the administration tothe human in need thereof a farnesyl protein transferase inhibitingamount of the compound of claim
 17. 25. A pharmaceutical compositioncomprising an effective amount of compound of claim 17 in combinationwith a pharmaceutically acceptable carrier.
 26. A method of treatingtumor cells in a human by inhibition of farnesyl protein transferasewherein the cells treated are pancreatic tumor cells, lung cancer cells,myeloid leukemia tumor cells, thyroid follicular tumor cells,myelodysplastic tumor cells, epidermal carcinoma tumor cells, bladdercarcinoma tumor cells, colon tumors cells, breast tumor cells orprostate tumor cells comprising administering to the human in needthereof a farnesyl protein transferase inhibiting effective amount of acompound of claim
 18. 27. A method of inhibiting farnesyl proteintransferase in a human comprising the administration to the human inneed thereof a farnesyl protein transferase inhibiting amount of thecompound of claim
 18. 28. A pharmaceutical composition comprising aneffective amount of compound of claim 18 in combination with apharmaceutically acceptable carrier.
 29. A method of treating tumorcells in a human by inhibition of farnesyl protein transferase whereinthe cells are pancreatic tumor cells, lung cancer cells, myeloidleukemia tumor cells, thyroid follicular tumor cells, myelodysplastictumor cells, epidermal carcinoma tumor cells, bladder carcinoma tumorcells, colon tumors cells, breast tumor cells or prostate tumor cellscomprising administering to the human in need thereof a farnesyl proteintransferase inhibiting amount of a compound of claim
 19. 30. A method ofinhibiting farnesyl protein transferase in a human comprising theadministration to the human in need thereof a farnesyl proteintransferase inhibiting amount of the compound of claim
 19. 31. Apharmaceutical composition comprising an effective amount of compound ofclaim 19 in combination with a pharmaceutically acceptable carrier.